We are studying the detailed mechanism of generation of "irreversibly sickled cells" (ISC) during incubation of deoxygenated sickle cells. The effects of intra and extracorpuscular agents are to be evaluated (e.g., ATP, DPG calcium, lysolecithin, prostaglandins). The effects of treatment with cyanate, nitrogen mustard, and membrane active agents on in vitro ISC generation is to be studied. We plan to investigate the possibility of developing suitable techniques for extra-corporeal treatment of red cells with agents such as cyanate using a continuous flow system such as the blood cell separator. We will measure the effects of intracellular polymerization of deoxy HbS on the environment of - SH and phosphate groups in the hemoglobin molecule using infrared spectroscopy. We will calculate the energy of deoxy HbS polymerization by measuring the increase in sickle blood O2 affinity produced by inhibition of sickling (nitrogen mustards).